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慢性暴露于骨形态发生蛋白-2 有利于单层培养扩增的人关节软骨细胞的软骨形成表达。

Chronic exposure of bone morphogenetic protein-2 favors chondrogenic expression in human articular chondrocytes amplified in monolayer cultures.

机构信息

Université de Lyon, Lyon F-69003, France.

出版信息

J Cell Biochem. 2010 Dec 15;111(6):1642-51. doi: 10.1002/jcb.22897.

Abstract

Articular cartilage is a specialized connective tissue containing chondrocytes embedded in a network of extracellular macromolecules such as type II collagen and presents poor capacity to self-repair. Autologous chondrocyte transplantation (ACT) is worldwide used for treatment of focal damage to articular cartilage. However, dedifferentiation of chondrocytes occurs during the long term culture necessary for mass cell production. The aim of this study was to investigate if addition of bone morphogenetic protein (BMP)-2, a strong inducer of chondrogenic expression, to human chondrocytes immediately after their isolation from cartilage, could help to maintain their chondrogenic phenotype in long-term culture conditions. Human articular chondrocytes were cultured according to the procedure used for ACT. Real-time PCR and Western blotting were performed to evaluate the cellular phenotype. Exogenous BMP-2 dramatically improves the chondrogenic character of knee articular chondrocytes amplified over two passages, as assessed by the BMP-2 stimulation on type II procollagen expression and synthesis. This study reveals that BMP-2 could potentially serve as a therapeutic agent for supporting the chondrogenic phenotype of human articular chondrocytes expanded in the conditions generally used for ACT.

摘要

关节软骨是一种特殊的结缔组织,其中包含嵌入细胞外大分子网络(如 II 型胶原)中的软骨细胞,自我修复能力差。自体软骨细胞移植(ACT)被广泛用于治疗关节软骨的局灶性损伤。然而,软骨细胞在大规模细胞生产所需的长期培养过程中会发生去分化。本研究旨在探讨在从软骨中分离出来后立即向人软骨细胞中添加骨形态发生蛋白(BMP)-2,一种强烈诱导软骨形成表达的物质,是否有助于在长期培养条件下维持其软骨形成表型。根据 ACT 中使用的程序培养人关节软骨细胞。通过实时 PCR 和 Western blot 评估细胞表型。外源性 BMP-2 通过刺激 II 型前胶原表达和合成,极大地改善了经过两个传代扩增的膝关节关节软骨的软骨形成特性。这项研究表明,BMP-2 可能作为一种治疗剂,用于支持 ACT 中常用条件下扩增的人关节软骨细胞的软骨形成表型。

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